STX 5对肝细胞癌转移的影响及其机制  

Effect of STX 5 on the metastasis of hepatocellular carcinoma and its mechanism

作　　者：郭婷婷 赵梓吟 何明阳 吴天松 徐斌 张斌[1] 吴泽华 韩冰[1] GUO Tingting;ZHAO Ziyin;HE Mingyang;WU Tiansong;XU Bin;ZHANG Bin;WU Zehua;HAN Bing(Department of Hepatobiliary and Pancreatic Surgery,The Affiliated Hospital of Qingdao University,Qingdao 266003,China)

机构地区：[1]青岛大学附属医院肝胆胰外科,山东青岛266003

出　　处：《精准医学杂志》2024年第3期236-241,246,共7页Journal of Precision Medicine

基　　金：山东省自然科学基金项目(ZR2020MH217)。

摘　　要：目的探讨STX5对肝细胞癌(HCC)转移的影响及其机制。方法收集2015年1—12月我院确诊为HCC的36例患者的临床资料,分析肿瘤组织中STX5表达水平与HCC患者临床病理特征的相关性。将人肝癌细胞MHCC97H分为A、B组,分别转染阴性对照质粒、过表达STX5质粒,将人肝癌细胞Huh7分为C、D组,分别转染阴性对照慢病毒、STX5敲减慢病毒,采用Western blot实验检测A~D组细胞内STX5蛋白表达水平,采用划痕实验检测A~D组细胞的迁移能力,采用Transwell实验检测A~D组细胞的迁移能力。对A、B组细胞经转录组学测序分析获得的差异基因进行GO功能和KEGG信号通路富集分析,实时荧光定量聚合酶链反应(RT-qPCR)检测A、B组细胞最显著差异表达基因的水平。将MHCC97H细胞分为E~H组,分别转染阴性对照质粒、过表达STX5质粒、阴性对照质粒+Sarilumab、过表达STX5质粒+Sarilumab,采用划痕实验检测E~H组细胞的迁移能力,采用Transwell实验检测E~H组细胞的迁移能力。结果肿瘤组织中STX5表达水平与患者的BMI、有无乙型肝炎病毒感染、肿瘤数量有关(P<0.05)。Western blot检测结果显示,B组与A组、C组与D组比较,细胞中STX5的表达显著增高(t=48.86、31.09,P<0.05)。Transwell实验和划痕实验结果显示,B组与A组、C组与D组比较,细胞迁移能力和划痕愈合百分比显著升高(t=7.95~31.09,P<0.05)。GO和KEGG富集分析显示,A、B组细胞差异基因主要富集在细胞迁移、炎症等相关功能和通路上;RT-qPCR实验结果显示,B组细胞中IL-6 mRNA的表达水平显著高于A组细胞(t=23.69,P<0.05)。Transwell和划痕实验结果显示,G组和E组、H组和F组比较,细胞迁移能力和划痕愈合百分比均显著降低(t=2.94~24.39,P<0.05)。结论STX 5可能通过上调IL-6 mRNA表达来促进肝细胞癌细胞的转移。Objective To investigate the effect of STX5 on the metastasis of hepatocellular carcinoma(HCC)and its mechanism.Methods Clinical data were collected from 36 patients who were diagnosed with HCC in our hospital from January to December 2015,and the correlation between the expression level of STX5 in tumor tissue and the clinicopathological features of HCC patients was analyzed.Human hepatoma cells MHCC97H were divided into groups A and B and were transfected with negative control plasmid and STX5 overexpression plasmid,respectively.Human hepatoma cells Huh7 were divided into groups C and D and were transfected with negative control lentivirus and STX5 knockdown virus,respectively.Western blot was used to measure the protein expression level of STX5 in groups A,B,C,and D,wound healing was used to measure the migration ability of cells in groups A,B,C,and D,and Transwell assay was used to measure the migration ability of cells in groups A,B,C,and D.Gene ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis were performed for the differentially expressed genes in groups A and B obtained by transcriptomic sequencing,and RT-qPCR was used to measure the levels of the most significantly differentially expressed genes in group A and B.MHCC97H cells were divided into groups E,F,G,and H and were transfected with negative control plasmid,STX5 overexpression plasmid,negative control plasmid+Sarilumab,and STX5 overexpression plasmid+Sarilumab,respectively.Scratch assay was used to measure the migration ability of cells in groups E,F,G,and H,and Transwell assay was used to measure the migration ability of cells in groups E,F,G,and H.Results The expression level of STX5 in tumor tissue was associated body mass index,presence or absence of hepatitis B virus infection,and the number of tumors(P<0.05).Western blot showed that group B had a significantly higher expression level of STX5 than group A,and group C had a significantly higher expression level of S

关 键 词：Qa-SNARE蛋白质类 白细胞介素6 癌 肝细胞 基因表达调控 肿瘤 肿瘤浸润 肿瘤转移 

分 类 号：R735.7[医药卫生—肿瘤] R730.261[医药卫生—临床医学]